抗人EGFRvⅢex单克隆抗体的制备及初步鉴定  

Preparation and characterization of monoclonal antibody against human EGFRvⅢex

作　　者：江秀玲[1] 胡有根[2] 石必枝[3] 

机构地区：[1]扬州职业大学,江苏扬州225000 [2]武警江苏总队医院泌尿外科 [3]上海市肿瘤研究所,癌基因及相关基因国家重点实验室,200000

出　　处：《实用医药杂志》2013年第9期810-813,共4页Practical Journal of Medicine & Pharmacy

摘　　要：目的研制抗人表皮生长因子受体突变体Ⅲ胞外区(EGFRvⅢex)单克隆抗体(mAb),并鉴定其特性。方法采用原核表达的EGFRvⅢex蛋白免疫BALB/c小鼠,以间接ELISA法筛选分泌特异性mAb的杂交瘤细胞,采用蛋白印迹、间接免疫荧光和免疫组化染色鉴定mAb的特异性。结果获得1株可稳定分泌mAb的杂交瘤细胞株。mAb的Ig亚类为IgM,效价为10-6。mAb可识别U87细胞膜上稳定转染的人EGFRvⅢex和SPC-A1细胞膜上高表达的EGFR。结论成功制备1株抗EGFRvⅢex的mAb,经蛋白印迹、免疫荧光和免疫组化染色检测,mAb的特异性良好,能够检测细胞上的EGFR和EGFRvⅢex,为肿瘤的治疗提供了良好的手段。Objective To study the preparation and characterization of monoclonal antibody against human. Methods BALB/c mice were immunized with prokaryotically expressed human EGFRv m ex protein. The mAb- positive hybridoma cells were screened by indirect ELISA. Specificity of mAb was characterized by Western blot, immunofluorescent staining and immunohischemical staining. Results One hybridoma cells secreting mAb was established. The isotype of mAb was IgM. The titer was 10-6. The mAb recognized human EGFRvUlex protein on EGFRvⅢ ex-transfected U87 cells and EGFR protein on SPC-A1 cells. Conclusion One anti-EGFRv lll ex mAb has obtained. The characterization of the mAb indicated the fine specificity of mAb by Western blot, immunofluorescent staining and immunohischemical staining, which can provided a powerful method for the tumor therapy.

关 键 词：EGFRvⅢex 单克隆抗体 鉴定 

分 类 号：Q71[生物学—分子生物学]